近日,西南大學于士將老師,使用IPHASE品牌產(chǎn)品:小鼠肝微粒體在《Journal of Hazardous Materials》全威期刊上發(fā)表文章《A down-regulated cytochrome P450 in Neoseiulus barkeri Hughes (Acari: Phytoseiidae) can dechlorinate and hydroxylate chlorpyrifos without producing chlorpyrifos-oxon》,影響因子12.2!
在有害昆蟲生物防治中農(nóng)業(yè)系統(tǒng)中存在的天敵資源發(fā)揮著重要作用,常見的有機磷類殺蟲劑如毒死蜱等在農(nóng)田噴灑作業(yè)時,對天敵的生存造成了負面影響,選育抗藥性天敵品系有助于生物防治和化學防治工作的協(xié)同開展。此前西南大學實驗室選育了一種捕食性天敵-巴氏新小綏螨毒死蜱中抗品系,在本研究中發(fā)現(xiàn)其抗性形成可能與P450解毒酶對毒死蜱代謝能力的改變有關。通過桿狀病毒蛋白表達系統(tǒng),獲得了巴氏新小綏螨三種P450微粒體蛋白,其活性與陽性對照鼠肝微粒體P450活性類似,CO差光譜顯示在450nm處具有吸收峰,利用NADPH再生系統(tǒng),建立了三種P450微粒體蛋白對毒死蜱體外代謝的反應體系,HPLC結合MS/MS分析發(fā)現(xiàn),一個在抗性品系中下調表達的P450可以將毒死蜱吡啶環(huán)5號位的氯離子脫去并羥基化,而該蛋白下調表達產(chǎn)生抗性的原因我們推測其代謝產(chǎn)物為毒死蜱的毒性激活物,減少該成分產(chǎn)生有助于減緩其對作用靶標的蛋白活性抑制。
摘要
Selection of chemical-resistant predatory mites is a good alternative to balance the contradiction between chemical control and biological control. Previously, a resistant strain of Neoseiulus barkeri for chlorpyrifos was obtained. In the current study, two up-regulated (NbCYP3A6, NbCYP3A16) and one down-regulated (NbCYP3A24) P450s were screened through differential expression analysis and other detoxification-related genes such as CCEs, GST, etc. were not found. 3D modelling and molecular docking indicated that the chlorine at position 5 on the pyridine ring of chlorpyrifos, as well as a methyl group, were closest to the heme iron of the enzymes (less than 5 Å). Three active recombinant P450 proteins were heterologously expressed and metabolized with chlorpyrifos in vitro. HPLC assay showed that only NbCYP3A24 could metabolize chlorpyrifos, with a metabolism rate of 21.60 %. Analysis of the m/z of metabolites by LC-MS/MS showed that chlorine at the 5C position of chlorpyrifos was stripped and hydroxylated, whereas chlorpyrifos-oxon, a common product of oxidation by P450, was not found. Knockdown of the NbCYP3A24 gene in the susceptiblestrain did reduce the.
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